ELISA kit for detecting food allergen based on IgD antibody

A technology for food allergy and antibody detection, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of difficult analysis, high detection cost, slow detection speed, etc., to achieve strong detection sensitivity, wide application range, improved The effect of sensitivity

Inactive Publication Date: 2018-11-23
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Detect and identify allergens in food by analyzing the characteristics of allergen proteins, but special large-scale precision instruments are required, the detection cost is high, and the analysis is difficult. Special testing personnel and analysts are required.
[0005] To sum up, although there are many methods for the detection of food allergens currently studied, they all have the problems of slow detection speed, high cost, and are not suitable for batch detection, and require specific analytical instruments for machine-reading, which restricts the detection of food allergens. Development of original assays
Most importantly, no food allergen-specific IgD test kits currently exist

Method used

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  • ELISA kit for detecting food allergen based on IgD antibody
  • ELISA kit for detecting food allergen based on IgD antibody
  • ELISA kit for detecting food allergen based on IgD antibody

Examples

Experimental program
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preparation example Construction

[0049] In the present invention, the preparation method of the detection plate coated with food allergen preferably comprises the following steps:

[0050] 1) diluting the food allergen with a coating buffer to obtain a food allergen coating solution;

[0051] 2) Add the food allergen coating solution obtained in the step 1) to the reaction well of the detection plate, overnight at 4°C;

[0052] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.

[0053] In the invention, the food allergen is diluted with the coating buffer solution to obtain the coating solution. In the present invention, the dilution method is not particularly limited, and a dilution technique known to those skilled in the art can be used. In the present invention, the coating buffer is preferably carbonate buffer. The concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L. The pH value of the carbonate ...

Embodiment 1

[0088] Coating: Dilute egg white, wheat and milk food allergens with carbonate buffer solution to a food allergen coating solution with a total protein concentration of 10 μg / ml; dilute human IgD protein with carbonate buffer solution to obtain a standard Coating solution (see Table 1 for protein concentration), add 0.05ml of coating solution to each reaction well of the polystyrene plate, and overnight at 4°C. On the next day, discard the solution in the wells, add 0.35ml washing buffer to each well, wash 3 times, and incubate for 30s each time (referred to as washing, the same below);

[0089]Blocking: Add 0.05ml of PBS buffer (blocking solution) containing 2% bovine serum albumin to each reaction well, pH: 7.2, incubate at 15-35°C for 120min, discard the solution in the well, and wash as above;

[0090] Add samples: sera from patients with suspected egg allergy, serum from patients with wheat allergy, serum from patients with milk allergy (serum to be tested), and normal hu...

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Abstract

The invention provides an ELISA kit for detecting a food allergen based on an IgD antibody, which belongs to the technical field of food allergen. The kit comprises the following reagents: a detectionplate coated with food allergen; a washing buffering solution; a sample diluting solution; a standard product or a positive quality control product; a biotin-marked anti-human IgD antibody preparation; an enzyme-labeled avidin solution; a substrate developing solution; and a terminating solution. The IgD is first used for detecting the food allergen; the ELISA kit prepared in an indirect method has the characteristics of high detection sensitivity, high accuracy and good repetition performance; and moreover, a great amount of samples can be treated at one time. Embodiments prove that the detection sensitivity of the kit provided by the invention reaches 2ng/ml.

Description

technical field [0001] The invention relates to the technical field of food allergen detection, in particular to a kit for detecting food allergens based on IgD antibodies, that is, a food allergen-specific IgD detection kit. Background technique [0002] The incidence of allergic diseases accounts for more than 30% of the world's total population, and is listed by the World Health Organization as one of the four major non-infectious diseases in the 21st century. In recent years, the incidence of allergic diseases has increased significantly and has become a public health issue of global concern. The epidemiological survey of allergic diseases by Guo Jia et al. shows that 22% of the total population of 1.2 billion suffer from allergic diseases. For example, allergic rhinitis, asthma, conjunctivitis, eczema, food allergy, drug allergy and severe anaphylaxis, etc., seriously affect people's quality of life and life safety. [0003] Food allergens such as eggs, peanuts, milk, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/535G01N33/543G01N33/577
CPCG01N33/6893G01N33/535G01N33/543G01N33/577G01N2333/4712
Inventor 何韶衡何萍
Owner 何韶衡
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